Stone S, Sacher M, Mao Y, Carr C, Lyons P, Quinn A M, Ferro-Novick S
Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Mol Biol Cell. 1997 Jul;8(7):1175-81. doi: 10.1091/mbc.8.7.1175.
Bet1p is a type II membrane protein that is required for vesicular transport between the endoplasmic reticulum and Golgi complex in the yeast Saccharomyces cerevisiae. A domain of Bet1p, that shows potential to be involved in a coiled-coil interaction, is homologous to a region of the neuronal protein SNAP-25. Here, we used in vitro binding studies to demonstrate that Bet1p plays a role in potentiating soluble NSF attachment protein receptor (SNARE) interactions. Mutational analysis points to the coiled-coil region as necessary for Bet1p function, and circular dichroism experiments support this theory. In vitro binding studies were also used to demonstrate that a direct interaction between Bet1p and Bos1p is required for the efficient interaction of the vesicle SNARE with its SNARE target. Genetic studies suggest that the interactions of Bet1p with Bos1p are regulated by the small GTP-binding protein Ypt1p.
Bet1p是一种II型膜蛋白，在酿酒酵母中，它是内质网与高尔基体复合体之间囊泡运输所必需的。Bet1p的一个结构域显示出参与卷曲螺旋相互作用的潜力，该结构域与神经元蛋白SNAP - 25的一个区域同源。在这里，我们通过体外结合研究证明，Bet1p在增强可溶性NSF附着蛋白受体（SNARE）相互作用中发挥作用。突变分析表明卷曲螺旋区域是Bet1p功能所必需的，圆二色性实验支持这一理论。体外结合研究还用于证明，Bet1p与Bos1p之间的直接相互作用是囊泡SNARE与其SNARE靶点有效相互作用所必需的。遗传学研究表明，Bet1p与Bos1p的相互作用受小GTP结合蛋白Ypt1p的调节。
